Objective To explore the effect and preliminary mechanism of the plant-derived immunostimulatory molecule,ophiopogonin,on enhancing the immune response of a subunit vaccine with the receptor-binding domain(RBD)of coronavirus spike protein as the antigen.Methods CCK-8 assay was used to determine the cytotoxicity of ophiopogonin D'(OPD')on bone marrow-derived dendritic cells(BMDCs).Female Balb/c mice were randomly divided into RBD,RBD/OPD',RBD/Alum,and control groups.The immunization dose was 5 μg of antigen per mouse and 100 μg of adjuvant per mouse,and immunization was carried out according to the intramuscular injection immunization procedure on days 0,21,and 42.The titers of specific IgG and its subtype antibodies were detected by ELISA.The cytokine levels in the supernatant of splenocytes were detected using ELISA.The number of splenocytes secreting IFN-γ was detected by ELISpot.Laser confocal microscopy was employed to observe the uptake of antigen by BMDCs.The phagocytic ability of BMDCs for antigen was quantitatively analyzed by flow cytometry.The mechanism of its enhanced immune effect was preliminarily explored using transcriptomics technology combined with bioinformatics research.Results When the concentration of OPD'was less than 5 μg/mL,the survival rate of BMDCs was 100%.After a single intramuscular injection in mice,except for a slight decrease in body weight,the other biochemical indicators were within corresponding normal ranges.After intramuscular injection immunization of the vaccine,the titers of serum-specific IgG,IgG1,and IgG2a in the RBD/OPD'group were significantly higher than those in the RBD group(P<0.05).Compared with the RBD group,the RBD/OPD'group induced a high-level Th1 cell immune response of IL-1β,TNF-α,and IFN-γ(P<0.01)and had more lymphocytes secreting IFN-γ(P<0.001).Laser confocal microscopy displayed that BMDCs took up more antigens after OPD'treatment,which was further confirmed with flow cytometry in quantitative analysis on antigen uptake rate(P<0.01).Transcriptomics results indicated that there was more significant enrichment of the PPAR signaling pathway in the RBD/OPD'group than the RBD group,suggesting that OPD'may activate the PPAR signaling pathway to exert its adjuvant effect.Conclusion OPD'effectively enhances the immune response of the RBD subunit vaccine,and its action mechanism may be related to the activation of the PPAR signaling pathway.

Objective:To investigate the role and underlying mechanism of parthanatos death in neonatal SD rats with bilirubin encephalopathy(BE).Methods:Eighty 3-day-old neonatal SD rats were selected and randomly divided into control group and BE group.The BE model was established by intraperitoneal injection of bilirubin solution,and the pathological changes in the hippocampus were observed by hematoxylin-eosin(HE)staining and Nissl staining.The protein expressions of the phosphorylation of the core histone protein H2AX(termed gamma H2AX),poly ADP-ribose polymerasw-1(PARP-1)and apoptosis-inducing factor(AIF)in hippocampus were detected by Western blot.Immuno-fluorescence staining was used to detect the expression and distribution of AIF in hippocampus.Results:Compared with the control group,neonatal SD rats developed jaundice 12 hours after bilirubin injection,accompanied by slow weight gain.HE staining and Nissl staining showed that the hippocampus in BE group were damaged and the content of Nissl bodies was decreased.Western blot results showed that the expression of γ-H2AX protein in hippocampus began to increase at 72 h after modeling(P＜0.05),and the levels of PARP-1 and AIF protein in hippocampus increased signif-icantly at 72 h after modeling(P＜0.05).Immunofluorescence staining showed increased AIF expression and nuclear translocation.Conclusion:Intraperitoneal injection of bilirubin can induce DNA damage in hippocampal neurons of neonatal SD rats and activate the PARP-1/AIF pathway to cause parthanatos death of hippocampal neurons.

Objective:To investigate the role and underlying mechanism of parthanatos death in neonatal SD rats with bilirubin encephalopathy(BE).Methods:Eighty 3-day-old neonatal SD rats were selected and randomly divided into control group and BE group.The BE model was established by intraperitoneal injection of bilirubin solution,and the pathological changes in the hippocampus were observed by hematoxylin-eosin(HE)staining and Nissl staining.The protein expressions of the phosphorylation of the core histone protein H2AX(termed gamma H2AX),poly ADP-ribose polymerasw-1(PARP-1)and apoptosis-inducing factor(AIF)in hippocampus were detected by Western blot.Immuno-fluorescence staining was used to detect the expression and distribution of AIF in hippocampus.Results:Compared with the control group,neonatal SD rats developed jaundice 12 hours after bilirubin injection,accompanied by slow weight gain.HE staining and Nissl staining showed that the hippocampus in BE group were damaged and the content of Nissl bodies was decreased.Western blot results showed that the expression of γ-H2AX protein in hippocampus began to increase at 72 h after modeling(P＜0.05),and the levels of PARP-1 and AIF protein in hippocampus increased signif-icantly at 72 h after modeling(P＜0.05).Immunofluorescence staining showed increased AIF expression and nuclear translocation.Conclusion:Intraperitoneal injection of bilirubin can induce DNA damage in hippocampal neurons of neonatal SD rats and activate the PARP-1/AIF pathway to cause parthanatos death of hippocampal neurons.

Objective:To compare the efficacy of domestic and imported hemostatic clips in preventing delayed post-polypectomy bleeding (DPPB) after endoscopic resection of colorectal polyps ≥ 10 mm.Methods:Clinical data of 789 patients who underwent endoscopic resection of colorectal polyps (polyp diameter ≥10 mm) in Beijing Friendship Hospital, Capital Medical University from January 2018 to December 2019 were collected. The patients were divided into DPPB group ( n=15) and non-DPPB group ( n=774). Univariate and multivariate logistic regression models were used to analyze the influential factors for DPPB. The patients using one type of hemostatic clip were divided into the domestic hemostatic clip group ( n=499) and the imported hemostatic clip group ( n=208). The efficacy of hemostatic clips in preventing DPPB in the two groups was compared. Results:Among the 789 patients undergoing endoscopic resection of colorectal polyps, 1.9% (15/789) suffered from DPPB. Multivariate logistic regression analysis showed that pedunculated polyp was an independent risk factor for DPPB ( OR=6.621, 95% CI: 2.278-19.241, P=0.001), and closure of mucosal defect was an independent protective factor for DPPB ( OR=0.169,95% CI: 0.050-0.570, P=0.004). Regardless of physician experience, there was no significant difference between the domestic and imported hemostatic clip group in preventing DPPB after endoscopic resection of colorectal polyps ≥10 mm [experienced physicians: 1.8% (7/385) VS 0.6% (1/175), χ2=1.314, P=0.445; common physicians: 2.6% (3/114) VS 3.0% (1/33), χ2=0.010, P>0.999]. The domestic hemostatic clip group paid for less medical expenses than the imported hemostatic clip group (experienced physicians: 1 433.51±889.02 yuan VS 3 033.97±1 686.87 yuan, t<0.001 , P<0.001; common physicians: 1 181.58±815.29 yuan VS 3 303.46±1 690.43 yuan, t<0.001 ,P<0.001). Conclusion:Pedunculated polyp is an independent risk factor for DPPB after endoscopic resection of colorectal polyp larger than 10 mm, and clipping can significantly reduce the risk for DPPB. There is no significant difference in the prevention of DPPB between domestic and imported clips, but domestic clips compared with imported clips yield less medical burden, which are suitable for promotion to primary hospitals and major clinical centers.

Objective To investigate the correlation between serum levels of antiphospholipid antibody(aPL)(ACA-IgG,ACA-IgM,β2-GPI-IgG,β2-GPI-IgM),LAC,ds-DNA,and ANA and preterm labor with pre-maturity,and to analyze the prediction of preterm labor with the combination of age,week of gestation,history of delivery,and history of miscarriage,so as to provide references for the prevention and treatment of preterm la-bor and to promote eugenics.Methods Through a retrospective study design,43 pregnant women with preterm la-bor with preeclampsia diagnosed and treated at Guangdong Provincial People's Hospital from June 2018 to Decem-ber 2020 were collected as a case group,and 47 healthy pregnant women of the same period and similar gestational age were randomly selected as a control group.aPL(ACA-IgG,ACA-IgM,β2-GPI-IgG,β2-GPI-IgM)and ds-DNA were detected by enzyme immunoassay(ELISA)using an enzyme immunoassay instrument,lupus anticoagulant(LAC)in plasma was detected by coagulometer,and ANA was detected by indirect immunofluorescence using an immunofluorescence analyzer,and the application of SPSS 24.0 software was used to statistically analyze the gen-eral information and laboratory test data.the age of the patients was combined,gestational week,birth history,miscarriage history and other general information,logistic regression analysis was performed to find the indepen-dent influencing factors related to preterm labor;the analysis was performed by using the subjects'work charac-teristic curve(ROC curve)to determine the area under the ROC curve(AUC),the best predictive value,sensi-tivity and specificity,and to analyze the predictive value of preterm labor with preterm labor.Results In this study,the pregnant women in the group of pregnant women with preterm labor with preeclampsia were aged 27～40 years,with a mean age of(29.93±3.91)years,and the gestational weeks at the time of blood collection were 27-36 weeks,with a mean gestational week of(31.96±2.35)weeks,while the pregnant women in the healthy control group during the same time period were aged 25～40 years,with a mean age of(30.74±3.44)years,and the gestational weeks at the time of blood collection were 28～36 weeks,with a mean gestational week of(32.84±2.13)weeks.In the same period,healthy control group pregnant women were aged 25～40 years,with a mean age of(30.74±3.44)years,and were 28～36 weeks pregnant at the time of blood collection.The β2-GPI-IgM level of pregnant women in the case group with preterm labor was significantly higher than that of pregnant women in the healthy control group at the same time,with statistically significant differences(P<0.05),while the differences in the levels of β2-GPI-IgG,ds-DNA,and LAC between the two groups were not statistically significant(P>0.05).The analysis of the ROC curves showed that the AUC of β2-GPI-IgM was 0.642(P<0.05),which was the highest in the preterm group,and the AUC was 0.642(P<0.05).0.05),which was an independent influencing factor of preterm labor;age,gestational week,labor history,and miscarriage history could not be used as independent in-fluencing factors of preterm labor.Conclusion β2-GPI-IgM is associated with threatened preterm labor,it can be used as a predictor of threatened preterm labor,and has clinical utility in the monitoring of threatened preterm labor in pregnant women.

Background::Endoscopic ultrasound (EUS)-guided transmural drainage for pancreatic fluid collections (PFCs) has become the first-line treatment with quicker recovery and more minor injury compared with surgery and percutaneous drainage. The efficacy of stents implantation and drainage for different PFCs remains controversial, especially lumen-apposing metal stents (LAMS). This study aimed to compare the efficacy and safety of LAMS drainage for pancreatic pseudocysts (PPC) and walled-off necrosis (WON).Methods::A meta-analysis was performed for LAMS drainage for WON and PPC by systematically searching PubMed, Cochrane, and Embase databases from January 2010 to January 2020. From 2017 to 2019, 12 patients who were treated with LAMS drainage for PFCs in our medical center were also reviewed and included in this study.Results::Combining 11 copies of documents with the data from our medical center, a total of 585 patients with PFCs were enrolled in this meta-analysis, including 343 patients with WON and 242 with PPC. The technical success rate in WON is not significantly different from that of PPC ( P = 0.08 > 0.05). The clinical success of LAMS placement was achieved in 99% vs 89% in PPC and WON, respectively (RR = 0.92, 95% CI: 0.86-0.98, P = 0.01 < 0.05). The further intervention of direct endoscopic necrosectomy was required by 60% of patients in WON group. There was no significant difference in the incidence of adverse events, including infection, bleeding, stent migration and stent occlusion, after LAMS placement between WON and PPC. Conclusions::Endoscopic ultrasound-guided LAMS for PFCs are feasible, effective with preferable technical and clinical success rates. The clinical effect of LAMS on PPC is slightly better than that of WON, but its adverse reactions still need to be verified in a large-sample prospective study.

Background::Endoscopic ultrasound (EUS)-guided transmural drainage for pancreatic fluid collections (PFCs) has become the first-line treatment with quicker recovery and more minor injury compared with surgery and percutaneous drainage. The efficacy of stents implantation and drainage for different PFCs remains controversial, especially lumen-apposing metal stents (LAMS). This study aimed to compare the efficacy and safety of LAMS drainage for pancreatic pseudocysts (PPC) and walled-off necrosis (WON).Methods::A meta-analysis was performed for LAMS drainage for WON and PPC by systematically searching PubMed, Cochrane, and Embase databases from January 2010 to January 2020. From 2017 to 2019, 12 patients who were treated with LAMS drainage for PFCs in our medical center were also reviewed and included in this study.Results::Combining 11 copies of documents with the data from our medical center, a total of 585 patients with PFCs were enrolled in this meta-analysis, including 343 patients with WON and 242 with PPC. The technical success rate in WON is not significantly different from that of PPC ( P = 0.08 > 0.05). The clinical success of LAMS placement was achieved in 99% vs 89% in PPC and WON, respectively (RR = 0.92, 95% CI: 0.86-0.98, P = 0.01 < 0.05). The further intervention of direct endoscopic necrosectomy was required by 60% of patients in WON group. There was no significant difference in the incidence of adverse events, including infection, bleeding, stent migration and stent occlusion, after LAMS placement between WON and PPC. Conclusions::Endoscopic ultrasound-guided LAMS for PFCs are feasible, effective with preferable technical and clinical success rates. The clinical effect of LAMS on PPC is slightly better than that of WON, but its adverse reactions still need to be verified in a large-sample prospective study.